Fig. 4.

Fatty acylation of RFX3 regulates ciliary gene expression and cilia formation. (A) Western blot analysis shows comparable expression levels of WT RFX3 and the C544S mutant in the established stable cells. (B) qRT-PCR results show expression of ciliary genes in different stable cell lines expressing vector control, WT RFX3, and C544S mutant. (C) Representative images of cilia in different NIH 3T3 stable cell lines expressing vector control, WT RFX3, and C544S mutant. Cells were stained with anti-acetylated tubulin antibody (red), anti-RFX3 antibody (green), and DAPI (blue). (Scale bars: 20 μm.) (D) Quantification of the percentage of ciliated cells in different stable cell lines. At least 200 cells were analyzed for each cell line in three independent experiments. Values represent the average ± SEM of three experiments. *P < 0.05, **P < 0.01, ***P < 0.001 (different from control; Student’s t test).